Development and Validation of a SPME-GC-MS Method for In situ Passive Sampling of Root Volatiles from Glasshouse-Grown Broccoli Plants Undergoing Below-Ground Herbivory by Larvae of Cabbage Root Fly, Delia radicum L

• Published in: Phytochemical analysis Vol. 27; no. 6; pp. 375 - 393
• Main Authors: Deasy, William, Shepherd, Tom, Alexander, Colin J., Birch, A. Nicholas E., Evans, K. Andrew
• Format: Journal Article
• Language: English
• Published: England Blackwell Publishing Ltd 01.11.2016; Wiley Subscription Services, Inc
• Subjects: Animals; Brassica; Brassica - chemistry; Chemical ecology; Delia radicum; Diptera - growth & development; Gas Chromatography-Mass Spectrometry - methods; Herbivory; in situ root volatiles analysis; Larva - physiology; Plant Roots - chemistry; Solid Phase Microextraction - methods; SPME-GC-MS; Volatilization; Brassica; SPME-GC-MS; Delia radicum; Chemical ecology; in situ root volatiles analysis
• ISSN: 0958-0344; 1099-1565
• DOI: 10.1002/pca.2637

Introduction Research on plant root chemical ecology has benefited greatly from recent developments in analytical chemistry. Numerous reports document techniques for sampling root volatiles, although only a limited number describe in situ collection. Objectives To demonstrate a new method for non‐invasive in situ passive sampling using solid phase micro extraction (SPME), from the immediate vicinity of growing roots. Methods SPME fibres inserted into polyfluorotetrafluoroethylene (PTFE) sampling tubes located in situ which were either perforated, covered with stainless steel mesh or with microporous PTFE tubing, were used for non‐invasive sub‐surface sampling of root volatiles from glasshouse‐grown broccoli. Sampling methods were compared with above surface headspace collection using Tenax TA. The roots were either mechanically damaged or infested with Delia radicum larvae. Principal component analysis (PCA) was used to investigate the effect of damage on the composition of volatiles released by broccoli roots. Results Analyses by gas chromatography‐mass spectrometry (GC‐MS) with SPME and automated thermal desorption (ATD) confirmed that sulphur compounds, showing characteristic temporal emission patterns, were the principal volatiles released by roots following insect larval damage. Use of SPME with in situ perforated PTFE sampling tubes was the most robust method for out‐of‐lab sampling. Conclusion This study describes a new method for non‐invasive passive sampling of volatiles in situ from intact and insect damaged roots using SPME. The method is highly suitable for remote sampling and has potential for wide application in chemical ecology/root/soil research. Copyright © 2016 John Wiley & Sons, Ltd. A non‐invasive method is described for in situ passive sampling of volatiles below‐ground from roots of glasshouse‐grown plants using SPME fibres located in pre‐positioned sampling devices, consisting of perforated PTFE tubes or tubes covered with steel mesh. When both designs of sampling device were evaluated with broccoli plants, similar temporal changes were observed in the production of sulphur containing root volatiles before and after induction of root damage by larvae of cabbage root fly, Delia radicum.